Attempts to express fragments of TgNup503 or TgTom40, or to overexpress TgNup503, resulted in the localization a TgTOM40 fragment to the nuclear periphery. (A) A scheme showing the TgNup503 fragments attempted to test for localization. (B) Transient expression of five TgNup503 fragments seen in A C-terminally Ty tagged and immunofluorescence with anti-Ty (Pink) and the mitochondrial marker TgMys (green). (C) AlphaFold model for T. gondii Tom40. TgTom40 fragment 83–155 highlighted in green. (D) Transient expression of TgTom40_83–155 C-terminally Ty tagged fragment, and immunofluorescence with anti-Ty (green) and two nuclear markers the DNA staining DAPI (grey) and anti-TgENO2 (pink). Signal quantification for DAPI, anti-TgENO2, and TgTom40 fragment (anti-Ty) in a parasite cross-section. (E) Transient expression of TgTom40_83–155 C-terminally Ty-tagged fragment into a line with Myc-tagged TgNup302, and immunofluorescence with anti-Ty (green) and anti-Myc (pink) was performed using super-resolution microscopy. As a control, immunofluorescence with anti-TgCDPK1 (green) and anti-Myc (pink) was done. Signal quantification for TgNup302 and TgTom40 fragment (anti-Ty) or TgCDPK1 in a parasite cross-section.